Myoferlin Knockdown and Overexpression

MYOF was knocked down in NCI-N87 and MKN45 cells using lentiviral shRNA vectors (GenePharma Technology) designed to target the sequences 5′-GAAAGAGCTGTGCATTATAAA-3’ and 5′-GCTGTGGAGAAGAAGTTTAAC-3′ in the MYOF coding region. Stable knockdown cells were isolated by selecting for puromycin resistance.
To overexpress MYOF, human MYOF complementary DNA (cDNA) was amplified and inserted into a lentiviral vector; as a control, an empty lentiviral vector (GenePharma Technology) was prepared similarly. The vectors were introduced into HGC27 and SNU1 cells, and cells were selected for puromycin resistance 2 weeks before experiments performed as previously described (25 (link)).